Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Sognsvannsveien 20, P.O. Box 1057 Blindern, N-0316, Oslo, Norway.
Naunyn Schmiedebergs Arch Pharmacol. 2014 May;387(5):407-17. doi: 10.1007/s00210-013-0953-1. Epub 2014 Jan 15.
Recently, we showed C-type natriuretic peptide (CNP)-induced negative inotropic (NIR) and positive lusitropic response (LR) in failing rat heart. We wanted to study whether, and if so, how phosphodiesterases (PDEs) regulate CNP-induced cyclic 3',5'-guanosine monophosphate (cGMP) elevation and functional responses. Inotropic and lusitropic responses were measured in left ventricular muscle strips and cyclic nucleotide levels, PDE activity and phospholamban (PLB) and troponin I (TnI) phosphorylation were measured in ventricular cardiomyocytes from Wistar rats with heart failure 6 weeks after myocardial infarction. CNP-mediated increase in global cGMP was mainly regulated by PDE2, as reflected by a marked amplification of the cGMP increase during PDE2 inhibition and by a high PDE2 activity in cardiomyocytes. PDE3 inhibition, on the other hand, caused no significant cGMP increase by CNP. The functional consequences did not correspond to the changes of cGMP. PDE3 inhibition increased the potency of the CNP-induced NIR and LR, while PDE2 inhibition desensitized the CNP-induced NIR, but not LR. A role for PDE2 on the maximal LR and PDE5 on the maximal NIR to CNP was revealed in the presence of PDE3 inhibition. CNP increased PLB phosphorylation about 25- to 30-fold and tended to increase TnI phosphorylation about twofold. As a whole, CNP-induced functional responses were only modestly regulated by PDEs compared to the cAMP-mediated functional responses to β1-adrenoceptor stimulation, which are highly regulated by PDEs. There is a mismatch between the CNP-induced cGMP increase and functional responses. Global cGMP levels are mainly regulated by PDE2 after CNP stimulation, whereas the functional responses are modestly regulated by both PDE2 and PDE3, indicating cGMP compartmentation by PDEs affecting CNP-induced responses in failing hearts.
最近,我们发现 C 型利钠肽(CNP)可引起衰竭大鼠心脏的负性肌力(NIR)和正性变力(LR)反应。我们想研究磷酸二酯酶(PDEs)是否调节 CNP 诱导的环 3',5'-鸟苷酸(cGMP)升高和功能反应,如果是,是如何调节的。左心室肌条测定心肌收缩力和心肌舒张功能,用免疫印迹法测定梗塞 6 周后心力衰竭大鼠心室肌细胞 cGMP 水平、PDE 活性和肌球蛋白轻链磷酸化(PLB)和肌钙蛋白 I(TnI)磷酸化。CNP 介导的 cGMP 整体增加主要受 PDE2 调节,这反映在 PDE2 抑制时 cGMP 增加明显放大,以及心肌细胞中 PDE2 活性较高。另一方面,PDE3 抑制不会引起 CNP 引起的 cGMP 增加。功能变化与 cGMP 的变化不一致。PDE3 抑制增加了 CNP 诱导的 NIR 和 LR 的效力,而 PDE2 抑制使 CNP 诱导的 NIR 脱敏,但不使 LR 脱敏。在 PDE3 抑制存在的情况下,揭示了 PDE2 在最大 LR 上的作用和 PDE5 在最大 NIR 上的作用。CNP 使 PLB 磷酸化增加约 25-30 倍,使 TnI 磷酸化增加约两倍。总的来说,与 PDE 高度调节的β1-肾上腺素能受体刺激引起的 cAMP 介导的功能反应相比,CNP 诱导的功能反应仅受到 PDE 的适度调节。CNP 诱导的 cGMP 增加与功能反应之间存在不匹配。CNP 刺激后,cGMP 水平主要由 PDE2 调节,而功能反应由 PDE2 和 PDE3 适度调节,表明 PDE 对 cGMP 进行区室化,影响衰竭心脏中的 CNP 诱导反应。